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完整蛋白质复合物的化学交联和质谱分析研究多亚基蛋白质组合体的体系结构
JoVE Journal
Biologia
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JoVE Journal Biologia
Combining Chemical Cross-linking and Mass Spectrometry of Intact Protein Complexes to Study the Architecture of Multi-subunit Protein Assemblies
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完整蛋白质复合物的化学交联和质谱分析研究多亚基蛋白质组合体的体系结构

DOI:
10.3791/56747-v

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10:01 min

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November 28, 2017

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, , , , ,
1Interdisciplinary research center HALOmem,Martin Luther University Halle-Wittenberg, 2Department of Chemistry,Kings College London

Capítulos

  • 00:05Título
  • 00:51Mass Spectrometry-based Protein Identification
  • 04:35Mass Spectrometry of Intact Protein Complexes (Native Mass Spectrometry)
  • 06:17Chemical Cross-linking Coupled with Mass Spectrometry
  • 07:19Results: Chemical Cross-linking and Mass Spectrometry of Multi-subunit Protein Complexes
  • 09:14Conclusion

Summary

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蛋白质复合体的结构对其功能至关重要。结合各种质谱技术证明了研究其组装的强大功能。我们提供化学交联和本机质谱的协议, 并展示这些互补技术如何帮助阐明 multi-subunit 蛋白组件的结构。

Tags

Chemical Cross-linkingMass SpectrometryIntact Protein ComplexesMulti-subunit Protein AssembliesStructural InformationCompositionConnectivityStoichiometryProtein Complex PurificationElectrophoresisCoomassie StainingProtein Band ExcisionDisulfide ReductionCysteine AlkylationTrypsin DigestionPeptide Extraction

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